Fig. 5. A spatial map of the dynamics of 5-integrin or -actinin across the cell. ICM results for a confocal image time series of 5-GFP expressed in a CHO-B2 cell plated for 1 hour on 2 µg ml^–1 fibronectin (A), or for a two-photon microscopy image time series of -actinin-GFP on a CHO-B2 cell plated for 24 hours on 5 µg ml^–1 fibronectin (B), (C) and (D). Both image series were collected at 37°C. A temporal ICM analysis was performed on each of the highlighted regions of size 128², 64² or 32² pixels. Image stacks were with 100 frames at 0.111 µm (A) or 0.118 µm (C,D) pixel size and a frame interval of 5 seconds. The temporal correlation functions were fit to Equations 11, 12 and 13 and the fit with the best R² value was used. The circles represent the root mean square average diffusion distance from the center of the circle for a 10 minute period based on the measured average diffusion coefficient for each region. The vectors represent the mean translation distance and direction over a 10 minute period based on the measured velocities for regions exhibiting flowing integrin populations. The colored bars depict the proportion of immobile (green), flowing (yellow) and diffusing (cyan) integrin or -actinin within each region. For the 5-GFP image series some areas appear to be off of the cell (e.g. area 1). If this was the case the analysis was limited to the image frames where the region of interest was completely on the cell. Region 6 was too small for the directional correlation analysis. Correlation velocity mapping was done for small areas around a retracting microspike just below region 3 in Fig. 5B. The arrows show the direction of the flow component of the correlation function and the size of the arrow is proportional to the magnitude of the velocity in that area of the cell. (D) Correlation analysis of images 70-100 of the image time series showing diffusion of -actinin after adhesion disassembly. (E) Spatial-temporal correlation functions for -actinin in area 1 shown in Fig. 5C. Notice the center of the correlation function peak moves from the center of the axis in the direction shown by the arrow as things flow towards the upper left quadrant. Scale bars, 5 µm (A,C,D) or 10 µm (B).